Difference between revisions of "Part:BBa K1951003:Design"
 
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__NOTOC__
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===Sequence optimization===
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<partinfo>BBa_K1951003 short</partinfo>
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<partinfo>BBa_K1951003 SequenceAndFeatures</partinfo>
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Snapgene has been used to remove the forbidden sites by changing in the substitution which didn't change the amino acid. The sequence has been sythetised by IDT<ref>http://eu.idtdna.com/site</ref>.
  
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===Codon optimization for <i>Escherichia coli</i>. ===
  
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====Design Notes====
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We obtimised codon for <i>Escherichia coli</i>. Codon optimization is a technical used to <u>improve the protein expression</u> in living organism by increasing the translational efficiency of gene of interest [1-4, 6-13, 15-18, 20-28]. This biobrick codon optimised <u>increase the functionality of gene</u>. To process it, we use codon optimization IDT software<ref> https://eu.idtdna.com/CodonOpt </ref>.
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We added prefix and suffix subsequences (containing EcoRI, XbaI and SpeI, PstI restriction sites respectively).
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If you use this biobrick in <i>Escherichia coli</i>, you can be sure that the protein produced will be highly expressed and well solubilised.
  
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====Source====
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=== Prefix and suffix addition===
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Prefix and suffix subsequences (containing restriction site EcoRI, XbaI and SpeI PstI respectively) have been added by a SLIC method with the following oligos :
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{| class="wikitable"
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| desD slic forward 
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| cgctaaggatgatttctgGAATTCGCGGCCGCTTCTAGATGAGTTTAGCTGATGCA
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|-
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| desD slic reverse
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| ttgcccttttttgccggaCTGCAGCGGCCGCTACTAGTATTATTAACGCCCGGCTAA
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|}.
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===Sequence and features===
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<partinfo>BBa_K1951003 Sequence And Features </partinfo>
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===Source===
  
 
Gene reference and its chromosomal position have been found on the following website : http://strepdb.streptomyces.org.uk/cgi-bin/dc3.pl?accession=AL645882&serial=2769&width=900&start=3031362&end=3041362&iorm=map
 
Gene reference and its chromosomal position have been found on the following website : http://strepdb.streptomyces.org.uk/cgi-bin/dc3.pl?accession=AL645882&serial=2769&width=900&start=3031362&end=3041362&iorm=map

Latest revision as of 13:42, 17 October 2016

Sequence optimization

Snapgene has been used to remove the forbidden sites by changing in the substitution which didn't change the amino acid. The sequence has been sythetised by IDT[1].

Codon optimization for Escherichia coli.

We obtimised codon for Escherichia coli. Codon optimization is a technical used to improve the protein expression in living organism by increasing the translational efficiency of gene of interest [1-4, 6-13, 15-18, 20-28]. This biobrick codon optimised increase the functionality of gene. To process it, we use codon optimization IDT software[2]. If you use this biobrick in Escherichia coli, you can be sure that the protein produced will be highly expressed and well solubilised.

Prefix and suffix addition

Prefix and suffix subsequences (containing restriction site EcoRI, XbaI and SpeI PstI respectively) have been added by a SLIC method with the following oligos :

desD slic forward cgctaaggatgatttctgGAATTCGCGGCCGCTTCTAGATGAGTTTAGCTGATGCA
desD slic reverse ttgcccttttttgccggaCTGCAGCGGCCGCTACTAGTATTATTAACGCCCGGCTAA
.

Sequence and features

BBa_K1951003 Sequence And Features Not understood

Source

Gene reference and its chromosomal position have been found on the following website : http://strepdb.streptomyces.org.uk/cgi-bin/dc3.pl?accession=AL645882&serial=2769&width=900&start=3031362&end=3041362&iorm=map Start : 3038895 End : 3040682 The website bellows is the source of the DNA sequence we used. http://www.ncbi.nlm.nih.gov/nuccore/NC_003888.3

References

  1. ↑ http://eu.idtdna.com/site
  2. ↑ https://eu.idtdna.com/CodonOpt